Skip to main content
NCBI home page
Springer logoLink to Springer
. 2026 Jul 30;34(9):6275–6289. doi: 10.1007/s10787-026-02332-0

Atriplex portulacoides L. derived phytochemicals mitigate acetic acid induced colitis in rats via orchestrating Nrf2/Keap1 signalling and LncRNAs gene expression

Maha Hassan 1, Marwa M Mona 2, Marwa M Abd-Elsalam 3, Mona El-Aasr 1, Amal Kabbash 1, Ghada Attia 1,✉
PMCID: PMC13558346  PMID: 42530774

Abstract

Background

This study is the first to elucidate the mechanistic actions of Atriplex portulacoides L. methanolic extract (APME) as a natural treatment of acetic acid-induced ulcerative colitis on rat model.

Methods

Phytochemical metabolic profiling of APME was determined through the determination of total phenolic and flavonoid contents, alongside negative mode LC–ESI–MS/MS analysis. Furthermore, chromatographic and spectroscopic investigations were employed to fractionate and structurally characterize the metabolites present within APME fractions. The therapeutic potential of APME in UC was evaluated by integrating histological and immunohistochemical analyses. Quantification of oxidative stress, inflammation-related, and lncRNAs (FENDRR and Neat1) gene expression was established.

Results

APME was evaluated for its total phenolic content and total flavonoid content. LC–ESI–MS/MS analysis in negative ionization mode identified 35 bioactive metabolites within APME. Stigmasterol (1) and 20-Hydroxyecdysone (2) were isolated by column chromatography from the n-hexane and ethyl acetate fractions, respectively, and characterized using spectroscopic analysis. APME dose-dependently and significantly (p < 0.05) reduced the pro-inflammatory cytokine TNF-α, mitigated malondialdehyde levels, and restored total antioxidant capacity in colonic tissue compared to the untreated ulcerative group. APME`s treatments markedly upregulated the mRNA expression levels of Nrf2 and HO-1, while concurrently suppressing NF-κB mRNA expression. Also, it showed a regulatory effect on FENDRR and Neat1. Histological, immunohistochemical, and morphometric assessments further supported these findings, demonstrating substantial improvements in colonic architecture and cellular integrity.

Discussion

Notably, APME represents a potential antioxidant and anti-inflammatory therapeutic candidate for UC. It modulates FENDRR and Neat1, revealing promising targets for IBD therapy.

Supplementary Information

The online version contains supplementary material available at https://doi.org/10.1007/s10787-026-02332-0.

Keywords: Atriplex portulacoides L., LC–ESI–MS/MS, Ulcerative colitis, Nrf2/Keap1 signalling pathway, TNF-α, NF-Κb, LncRNAs

Introduction

Immune-mediated inflammatory diseases are diverse disorders marked by abnormal immune responses and dysregulated immunological pathways, affecting various body organs. Gastrointestinal IMIDs overlap, complicating diagnosis, including inflammatory bowel disease [IBD], coeliac, vasculitis, gastroenteritis, sarcoidosis, colitis, and microscopic colitis. IBD primarily consists of two distinct clinical entities: Crohn’s disease [CD] and ulcerative colitis [UC] (Vuyyuru et al. 2022).

The pathogenesis of IBD is complex and not yet fully understood. It involves multifactorial interplay among genetic susceptibility, environmental factors, immune system dysregulation, and disturbances in the composition and function of the gut microbiota (Zhang and Li 2014).

The core mechanism underlying IBD pathogenesis involves an abnormal immune response to the intestinal microbiota. In healthy individuals, the immune system maintains homeostasis by distinguishing between harmless commensal organisms and harmful pathogens. However, in those with genetic susceptibility, this regulatory balance is disrupted. Environmental factors—such as diet, infections, and stress—can intensify this imbalance, promoting an exaggerated inflammatory response within the gut (Abdulla and Mohammed 2022).

NF-κB is a key inducible transcription factor with critical roles in numerous physiological and pathological processes, making it a prominent target in therapeutic strategies, particularly in cancer and inflammatory diseases (Li et al. 2015; Oeckinghaus et al. 2011). Although NF-κB exhibits complex and multifaceted functions, it does not operate in isolation. Instead, it engages in extensive crosstalk with other signaling cascades to mediate a wide range of cellular responses (Attiq et al. 2018). Its activation can be triggered by diverse stimuli, including pro-inflammatory cytokines, oxidative stress, and microbial infections, leading to downstream effects on cell proliferation, differentiation, survival, immune modulation, and intercellular communication (Gerondakis et al. 1999; Mattson and Meffert 2006).

Experimental models, particularly in mice, have demonstrated the importance of Rel/NF-κB and IκB family members in regulating these pathways through genetic manipulation such as transgenic and knockout approaches (Courtois and Gilmore 2006). Disruption of NF-κB signaling has been implicated in the pathogenesis of a variety of disorders, including autoimmune and inflammatory diseases, metabolic syndromes, and cancers (Taniguchi and Karin 2018). Oxidative stress is a central feature of IBD pathophysiology, where reactive oxygen species [ROS] induce lipid peroxidation, protein oxidation, and DNA damage in intestinal epithelial cells. Several transcription factors, including nuclear factor erythroid 2-related factor 2 [Nrf2], heme oxygenase-1 [HO-1], and NF-κB, regulate redox homdamage in the immune system and inflammatory signaling. While Nrf2 and HO-1 exert protective roles by mitigating oxidative stress, Kelch-like ECH-associated protein 1 [Keap1] inhibits Nrf2 activity, exacerbating cellular injury (Li et al. 2015). Additionally, β-actin serves as a housekeeping gene in gene expression studies related to these pathways.

Approximately 30% of patients exhibit primary non-responsiveness to anti-TNFα therapy. Moreover, among initial responders, up to 10% experience a loss of therapeutic efficacy annually. Existing pharmacological options are also associated with considerable risks, including infections and malignancies. These limitations highlight a critical unmet need for the development and clinical integration of more effective therapeutic agents or combination regimens with improved safety profiles (Hazel and O'Connor 2020).

Plant-derived natural products have long been recognized for their beneficial effects on human health, historically utilized for purposes such as pain relief, wound healing, gastrointestinal support, and various other therapeutic applications (Davila and Papada 2023). In recent decades, these natural compounds have garnered significant scientific attention, prompting extensive research focused on the identification, isolation, quantification, and evaluation of their bioactive constituents and associated biomedical properties.

Atriplex portulacoides L. (AP) is a halophytic perennial shrub commonly known as sea purslane. It is widely distributed across African, European, and North American coasts, with limited presence along some Asian shores (Kubiak-Martens et al. 2015). It belongs to the family Amaranthaceae and is one of the eighteen species of the genus Atriplex found in Egypt. In vivo investigation has reported the therapeutic potential of some Atriplex species in the management of ulcerative colitis (Abd El Raheim et al. 2013).

AP is traditionally used as animal fodder and as a source of nutrition (Zanella and Vianello 2020). Hamada (2025) revealed the anti-cytogenotoxic effect of AP`s crude extract in mice bone marrow (Hamada et al. 2025).

Recent studies highlight long non-coding RNAs [lncRNAs] in IBD pathogenesis, particularly FENDRR [FOXF1 adjacent non-coding developmental regulatory RNA] and Neat1 [nuclear paraspeckle assembly transcript 1]. Neat1 drives cytokine expression, mitochondrial stress, and apoptosis, while FENDRR regulates macrophage activation, suggesting a biomarker for gastrointestinal tumorigenesis (He et al. 2019).

Herein, a phytochemical metabolic profiling of APME was accomplished via total phenolic, total flavonoid, and liquid chromatography- electrospray negative mode ionization tandem mass spectrometric [LC–ESI–MS/MS] analyses. Additionally, chromatographic and spectroscopic studies were conducted to separate and identify metabolites present in APME’s fractions. We evaluated the unrevealed therapeutic potential of APME in the treatment of UC via integrating histological and immunohistochemical assessments with quantifying the oxidative stress and inflammation-related gene expression. We also explored for the first time the modulation of FENDRR and Neat1 expressions by APME as potential molecular targets in IBD therapy.

Materials and methods

Plant material for APME chromatographic and biological studies

Aerial parts of A. portulacoides L. were collected from Kafr El-Sheikh Baltim coastal road, Egypt, in January 2019. The plant was kindly verified by Prof. Dr. Ahmed Sharaf eldin, Professor of Plant Ecology, Botany Department, Faculty of Science, Tanta University. Voucher sample No. PGAAP012019 was deposited at the herbarium of the Pharmacognosy department, Faculty of Pharmacy, Tanta University. The plant material was shade-dried, then ground into powder and kept in firmly closed containers.

2.8 kg of dried AP aerial parts powder were extracted through maceration (5 × 16 L) using methanol until complete exhaustion. The extract was then concentrated under reduced pressure using a rotary evaporator, yielding 120 g of APME for phytochemical and biological studies.

A total of 20 g of APME was suspended in 0.5% methyl cellulose to prepare dosing solutions corresponding to two final concentrations: 100 mg/kg and 200 mg/kg of rat body weight for use in the biological experiments.

Instruments and reagents for APME chromatographic and biological studies (Supplementary materials).

Total phenolics & total flavonoids examination of APME: (Supplementary materials).

LC–ESI–MS /MS Analysis of APME: (Supplementary materials).

Chromatographic separation and spectroscopic identification of metabolites from APME’s fractions: (Supplementary materials).

Materials for the APME biological study

Induction of Ulcerative Colitis: (Supplementary materials).

Experimental animals

The study was conducted using 50 healthy adult male albino rats, each weighing between 200 and 250 g. All animals were housed under controlled environmental conditions (12-h light–dark cycle; 24 ± 2 °C; 60–70% relative humidity) and acclimatized for two weeks before the initiation of the experiment.

Ethics statement

Animal handling was strictly adhered to the approved guidelines of the research ethics committee for the care and use of laboratory animals, at the Faculty of Pharmacy, Kafrelsheikh University (approval number of KFS-IACUC/301/2025). Following the ethical standards, the Egyptian guide for the care and use of laboratory animals has been used. The local ethics committee allowed the experimental protocol, which was fully committed to the ARRIVE criteria for reporting in vivo investigations and employing laboratory animals. The animals were humanely euthanised by administration of an intraperitoneal injection of sodium pentobarbital (50 mg/kg) at the end of the experiment.

Experimental design

The animals were randomly allocated into five groups, with 10 rats in each:

  • Group 1 (Control Group): This group was further subdivided into two subgroups (n = 5 each):

  • Subgroup I received the vehicle control (methyl cellulose) orally at a volume of 1 mL daily for 10 consecutive days. Also, the control group was given a single intra-rectal (IR) dose of 2 mL of 0.9% normal saline on the induction day.

  • Group 2 (APME 200 mg/kg): Rats were treated orally with 200 mg/kg of APME dissolved in 0.5% methyl cellulose for three days prior to colitis induction and continued for six additional days, including the day of induction. On the induction day, they also received a single intra-rectal instillation of 2 mL of 0.9% saline.

  • Group 3 (Acetic Acid Group): Ulcerative colitis was induced in this group by intra-rectal administration of 2 mL of 3% acetic acid in normal saline. No treatment was administered to this group.

  • Group 4 (APME 100 mg/kg + Acetic Acid): Rats received 100 mg/kg of APME orally for three days prior to acetic acid induction and continued for four days post-induction.

  • Group 5 (APME 200 mg/kg + Acetic Acid): This group received the higher dose of APME (200 mg/kg) following the same regimen as Group 4.

On the 10th day of the experiment, all animals were humanely euthanised under deep anaesthesia induced by an intraperitoneal injection of sodium pentobarbital (50 mg/kg). Colonic tissues were then carefully excised, rinsed with saline, and processed for subsequent histological and immunohistochemical evaluations, following established protocols for experimental colitis models (Owusu et al. 2020).

Histological examination

For histopathological evaluation, colonic tissues from all experimental groups were fixed in 10% neutral buffered formalin, dehydrated, and embedded in paraffin to prepare tissue blocks. Sections were cut at a thickness of 5 µm and processed for histological analysis. The sections were stained with hematoxylin and eosin (H&E) to assess general tissue morphology, inflammatory infiltration, epithelial damage, and structural integrity of the colonic mucosa, following standard histological procedures (Bancroft and Layton 2010; Millar et al. 1996).

Immunohistochemical Study: (Supplementary materials).

Morphometric Analysis: (Supplementary materials).

Quantitative Real-Time PCR (qRT-PCR): (Supplementary materials).

RNA Extraction: (Supplementary materials).

Primer Design and PCR Reaction: (Supplementary materials).

Gene Expression Analysis: (Supplementary materials).

Measurement of TNF-α: (Supplementary materials).

Assessment of Oxidative Stress Biomarkers in Colon Homogenates: (Supplementary materials).

Assessment of Gene Expression Levels of β–Actin, Nrf2, Keap1, NF-κB, and HO-1 mRNA: (Supplementary materials).

Assessment of Gene Expression Levels of Fendrr and Neat1 lncRNAs: (Supplementary materials).

Statistical analysis: (Supplementary materials).

Result

Phytochemical investigation of APME

Total phenolic and flavonoid contents

APME was found to contain 10.89 ± 0.95 mg GAE/g extract and 12.8 ± 0.84 mg RE/g extract of total phenolic and flavonoid compounds, respectively.

APME LC–ESI–MS/MS-based metabolic profiling

Tentative metabolic profiling of APME using negative mode LC–ESI–MS/MS identified 35 metabolites of several phytochemical classes, including organic acids, cinnamic acid derivatives, flavonoid aglycones and glycosides, biflavonoids, carbohydrates, purines, pyrimidines, amino acids, and volatile compounds (Table S1 and Figure S1). These results were in coherence with Liberian data and previously reported literatures.

Organic acids

Five deprotonated molecular ion peaks appeared at m/z 133.01425 (Abdl Aziz et al. 2024), 175.0612 (Sri Harsha et al. 2018), 165.05573 (Abdl Aziz et al. 2024), 137.02441, and 153.01933 (Collins 2022), referring to D-(+)-malic acid 1, 2-isopropylmalic acid 2, D-3-Phenyllactic acid 3, p-hydroxybenzoic acid 4, 2,5-dihydroxybenzoic acid 5, respectively.

Cinnamic acid derivatives

Two characteristic phenolic acids were recognized as caffeic acid 6 and rosmarinic acid 7 with [M-H] ¯ peak at m/z 179.03499 (Abdl Aziz et al. 2024) and 359.07724 (Stanojević et al. 2018), respectively.

Flavonoids and their glycosides

Two flavonol-type aglycones were detected as myricetin 8 at m/z 317.0303 and 3,5, 7-trihydroxy-4’-methoxyflavone 15 at m/z 299.05612 (Abdl Aziz et al. 2024). However, four flavonol -O-glycosides of variant glycone moieties were detected as kaempferol-3-O-α-L-rhamnoside 10, isorhamnetin-3-O-rutinoside 11, kaempferol-7-O-neohesperidoside 13, kaempferol-3-Glucuronide 14. These deprotonated molecular ion peaks appeared at m/z 431.09836 (Abdl Aziz et al. 2024), 623.16174 (Abdl Aziz et al. 2024), 593.15118, and 461.07254 (Fabre et al.2001; Yuan et al. 2008), respectively. Moreover, the flavone`s glycoside baicalein-7-O-glucuronide 9 was detected at m/z 445.07764 (Abdl Aziz et al. 2024), while the hydroxychalcone glycoside okanin-4’-O-glucoside 12 appeared at m/z 449.10895. The only detected biflavonoid at m/z 577.13513 was procyanidin B2 16.

Carbohydrates and sugar derivatives

Different carbohydrates were observed as the three sugar acids L-( +)-Tartrate 17, gluconate 18, and mucate 19, the sugar alcohol galactinol dihydrate 20, the disaccharide sucrose 21, and the oligosaccharides nystose trihydrate 22. Their deprotonated peaks were measured at m/z 149.00916, 195.05103 (Abdel Aziz et al. 2024), 209.0309 (El-Zawahry et al. 2025), 378.10895 (Abdelgawad et al. 2024), 341.10895 (Taylora et al. 2005), and 665.2783, respectively.

Purines and pyrimidines

The purine nucleosides inosine 23 and xanthosine 24 appeared at m/z 267.07349 and 283.06842 (Li et al. 2009), respectively. Other pyrimidine nucleosides were identified as, 2’-Deoxyuridine-5’-triphosphate sodium salt 25, uridine 5'-monophosphate 26, UDP-xylose 27, 2’-deoxyuridine-5’-triphosphate sodium salt uridine 5'-diphosphate 28, uridine 5'-diphosphate 29, thymidine-3’,5’ cyclic monophosphate sodium salt 30 and uridine 5’-diphosphoglucuronic acid 31, These pyrimidine derivatives showed [M-H] ¯ peaks values at m/z 307.0423, 323.1175, 535.03717, 466.966, 303.03876 and 579.02704, respectively.

Miscellaneous metabolites

The volatile compound ˠ-Terpinene 32 was identified at m/z 135.11792 (Ryhage and Sydow 1963), while the two amino acids L-5- oxoproline 33, trans-4-hydroxy-L-proline 34, and L-β-homoisoleucine 35 were detected at m/z 128.03532, 130.05096, and 144.04691, respectively.

Isolation and spectroscopic identification of components at AP fractions (supplementary material)

APME fractionation, chromatography-based purification, and spectral analysis (supplementary material) revealed the isolation of stigmasterol 1 (20 mg of an amorphous white powder) and 20-hydroxyecdysone 2 (8 mg of a yellowish amorphous solid) from its n-hexane and ethyl acetate fractions, respectively (Fig. 1).

Fig. 1.

Fig. 1

Open in a new tab

Compounds isolated from AP fractions

In Vivo evaluation of APME efficacy in UC Treatment

Guided by the high LD50 of an anti-ulcer activity of some species in the genus Atriplex (Abd El Raheim et al. 2013), no animal mortality was observed, indicating that APME was well-tolerated at the tested doses. The control group showed no significant differences in histopathological findings or biochemical outcomes. Therefore, they were collectively designated as the unified control group for subsequent analyses.

Similarly, animals in the APME-only treatment group (Group 2), which did not undergo colitis induction, showed no noticeable alterations in the histological architecture of the colon. Additionally, no significant differences were found between this group and the control group regarding all measured parameters. These results confirm that APME, when administered alone, exerted no adverse structural or inflammatory effects on colonic tissues, supporting its safety in vivo.

Light microscopic assessment of colonic tissue (H&E staining)

Histological evaluation of hematoxylin and eosin (H&E)-stained sections from the control group (G1) revealed intact colonic architecture characterized by well-organized, tightly packed simple tubular glands (crypts of Lieberkühn) extending into the muscularis mucosa. The crypts were lined by columnar absorptive epithelial cells displaying apical brush borders, basal oval nuclei, and numerous goblet cells with vacuolated cytoplasm. Occasional intraepithelial lymphocytes were also observed, reflecting normal mucosal immune surveillance (Fig. 2A. Similarly, the colonic sections of the APME-treated rats (G2) display preserved histological features with no evident alterations in crypt architecture or goblet cell distribution (Fig. 2B, supporting the non-toxic profile of APME.

Fig. 2.

Fig. 2

Open in a new tab

Hematoxylin and eosin (H&E)-stained sections of colonic mucosa from different experimental groups; A-F (magnification × 200, scale bar = 50 μm). A Group 1 (control) and B Group 2 (200 mg/kg APME) show normal mucosal architecture with numerous, regularly arranged, and closely packed crypts of Lieberkühn C extending through the mucosa and resting on the muscularis mucosa (Ms). The surface epithelium and crypt lining consist mainly of columnar epithelial cells (white arrowheads) with apical brush borders and basal nuclei, along with abundant goblet cells (arrows) displaying vacuolated cytoplasm and basal nuclei. Goblet cells increase in number toward the crypt base, while columnar cells become fewer. The lamina propria contains resident immune cells, mainly lymphocytes. C, D Group 3 (colitis-induced) exhibits complete crypt destruction (C), loss of surface epithelium (white arrowhead), absence of goblet cells, homogenous acidophilic areas in the lamina propria (black arrowhead), and extensive infiltration of inflammatory cells (star) reaching the muscularis mucosa. E Group 4 (100 mg/kg APME + colitis) shows focal crypt distortion, a marked reduction in goblet cells (white arrowheads), widened intercryptal spaces (line), homogenous acidophilic areas (black arrowhead), and mononuclear cell infiltration in the lamina propria (star). F Group 5 (200 mg/kg APME + colitis) reveals preserved mucosal architecture with intact crypts, continuous surface epithelium (white arrowhead), numerous goblet cells (arrows), minimal acidophilic change (black arrowhead), and mild mononuclear infiltration (star)

In contrast, H&E-stained sections from the ulcerative colitis group (G3) demonstrated extensive mucosal damage. The colonic crypts were severely disrupted, with a complete loss of surface epithelium, marked depletion or total absence of goblet cells, and areas of mucosal detachment and hemorrhage (Fig. 2C. Dense mononuclear cellular infiltrates were observed within the lamina propria, extending into the muscularis mucosa, reflecting active inflammation (Fig. 2D).

Colonic sections from the G4 group (100 mg/kg APME + A.A.) exhibited partial protection, with no frank ulceration but evidence of focal crypt architectural distortion and intercryptic space widening. A limited number of goblet cells were identified, and moderate mononuclear infiltration extended into the muscularis mucosa (Fig. 2E, indicating an intermediate anti-inflammatory response.

Notably, treatment with the higher dose of APME (G5; 200 mg/kg + A.A.) resulted in near-complete preservation of colonic architecture. Crypt integrity was restored, surface epithelium remained continuous, and goblet cell populations were well maintained. Only minimal inflammatory cell infiltration was detected in Fig. 2F, suggesting a robust histoprotective and anti-inflammatory effect of APME at this dosage.

Immunohistochemical analysis

TNF-α expression

Immunohistochemical staining for TNF-α revealed minimal expression in both the control group (G1) and the APME-only group (G2, 200 mg/kg), suggesting a baseline physiological level of this proinflammatory cytokine (Fig. 3(1)A&B). In contrast, colonic tissues from the ulcerative colitis group (G3) exhibited markedly intense TNF-α immunoreactivity, indicative of severe inflammation (Fig. 3(1)C).

Fig. 3.

Fig. 3

Open in a new tab

Immunohistochemical staining of inflammatory and stress markers [1 TNF-α, 2 P53, and 3 COX-2] in the colonic mucosa across experimental groups (A–E), and quantitative analysis of marker-positive cells (F) (magnification × 200; scale bar = 50 μm). 1 TNF-α expression; A Group 1 (control) and B Group 2 (200 mg/kg APME) display minimal TNF-α immunoreactivity (arrowheads). C Group 3 (ulcerative colitis) shows strong positive TNF-α expression (arrowheads), indicating a heightened inflammatory response. D Group 4 (100 mg/kg APME + colitis) exhibits moderate TNF-α immunoreactivity (arrowheads). E Group 5 (200 mg/kg APME + colitis) demonstrates reduced TNF-α expression, comparable to control levels (arrowheads). F Histogram showing the mean area percentage of TNF-α-positive cells in groups G1–G5. ^a^ P < 0.05 vs. Group 1; ^b^ P < 0.05 vs. Group 3; ^c^ P < 0.05 vs. Group 4. 2 P53 expression; A Group 1 (control) and B Group 2 (200 mg/kg APME) exhibit minimal nuclear P53 immunoreactivity (arrowheads). C Group 3 (ulcerative colitis) reveals strong P53 expression (arrowheads), suggesting increased cellular stress and damage. D Group 4 (100 mg/kg APME + colitis) shows moderate P53 immunoreactivity (arrowheads). E Group 5 (200 mg/kg APME + colitis) displays reduced P53 expression with minimal staining (arrowheads), approaching baseline levels. F Histogram depicting the mean area percentage of P53-positive cells across G1–G5. ^a^ P < 0.05 vs. Group 1; ^b^ P < 0.05 vs. Group 3; ^c^ P < 0.05 vs. Group 5. 3 COX-2 expression; A Group 1 (control) and B Group 2 (200 mg/kg APME) exhibit weak cytoplasmic COX-2 staining in a few epithelial cells (arrowheads). C Group 3 (Ulcerative colitis) shows intense COX-2 immunoreactivity in epithelial and lamina propria cells (arrowheads), indicating increased inflammatory activity. D Group 4 (100 mg/kg APME + colitis) reveals moderate COX-2 staining (arrowheads). E Group 5 (200 mg/kg APME + colitis) demonstrates COX-2 levels like the control (arrowheads), indicating inflammation resolution. F Histogram illustrating the mean area percentage of COX-2-positive cells in groupsG1–G5. ^a^ P < 0.05 vs. Group 1; ^b^ P < 0.05 vs. Group 3; ^c^ P < 0.05 vs. Group 5

Interestingly, animals in G4 (100 mg/kg APME + colitis) demonstrated moderate TNF-α staining (Fig. 3(1)D), indicating a partial attenuation of the inflammatory response. The most pronounced therapeutic effect was observed in G5 (200 mg/kg APME + colitis), where TNF-α expression was markedly reduced and comparable to the control Fig. 3(1)E, confirming the dose-dependent anti-inflammatory efficacy of APME.

p53 expression

Negligible p53 immunoreactivity was seen in both the control (G1) and APME-only (G2) groups, suggesting no genotoxic or apoptotic stress under normal conditions, Fig. 3(2)A&B. However, G3 (colitis group) exhibited intense p53 expression, consistent with epithelial DNA damage and stress-induced apoptotic signalling (Fig. 3(2)C).

Treatment with APME at 100 mg/kg (G4) resulted in a moderate decrease in p53 expression (Fig. 3(2)D), whereas the higher dose of 200 mg/kg (G5) restored p53 levels near baseline, like control levels (Fig. 3(2)E). This suggests that APME's antioxidant properties may mitigate oxidative DNA damage and inhibit apoptotic signalling via p53 downregulation.

COX-2 expression

Immunostaining for COX-2 in colon tissues from the control group (G1) and the APME-only group (G2; 200 mg/kg) demonstrated weak cytoplasmic immunoreactivity, localized mainly to scattered surface and glandular epithelial cells, reflecting basal expression levels of this inducible enzyme under non-inflammatory conditions Fig. 3(3)A&B.

In contrast, the ulcerative colitis group (G3), subjected to acetic acid instillation without treatment, displayed intense cytoplasmic COX-2 expression in both surface and glandular epithelium, consistent with severe mucosal inflammation and epithelial injury Fig. 3(3)C.

Interestingly, colonic sections from G4 (100 mg/kg APME + colitis) still exhibited elevated COX-2 immunoreactivity in numerous epithelial cells Fig. 3(3)D, suggesting that the lower dose of APME offered only partial anti-inflammatory protection. However, sections from G5 (200 mg/kg APME + colitis) revealed a marked reduction in COX-2 positive cells, Fig. 3(3)E, approaching control levels. This significant suppression highlights APME’s dose-dependent anti-inflammatory potential, likely mediated through attenuation of oxidative stress and modulation of the NF-κB/COX-2 axis.

Morphometric analysis and statistical evaluation

Quantitative morphometric evaluation of the immunohistochemical staining revealed a significant elevation (p < 0.05) in the mean percentage area of positive immunoreactivity for TNF-α, P53, and COX-2 in the colitis-induced group (G3) when compared to the control group (G1). This sharp increase reflects the heightened inflammatory and apoptotic activity associated with acetic acid-induced ulcerative colitis.

Conversely, treatment with APME demonstrated a dose-dependent ameliorative effect. Both the low-dose treatment group (G4, 100 mg/kg b.w.) and the high-dose treatment group (G5, 200 mg/kg b.w.) showed a statistically significant reduction (p < 0.05) in TNF-α, P53, and COX-2 expression levels when compared to the colitis group (G3). Furthermore, a notable difference was observed between the two treated groups, with G5 exhibiting a more pronounced decrease in the expression of these inflammatory and apoptotic markers than G4.

These findings, supported by the graphical data presented in Fig. 3(1F, 2F, and 3F), underscore the potential of APME to downregulate inflammation and oxidative stress-induced cellular damage in a dose-responsive manner.

Quantitative evaluation of APME’s anti-inflammatory and antioxidant effects in experimental UC

The ulcerative colitis group (G3) demonstrated a significant increase in TNF-α gene expression (Fig. 4a) compared to the healthy control (G1), confirming the occurrence of acute mucosal inflammation. Conversely, treatment with APME resulted in a significant suppression of TNF-α expression (p < 0.05) in a dose-dependent manner compared to the ulcerative colitis group G3, with the higher dose showing a more pronounced anti-inflammatory response.

Fig. 4.

Fig. 4

Open in a new tab

Assessment of proinflammatory and oxidative stress biomarkers a TNF-α via ELISA, b Malondialdehyde (MDA) and c Total antioxidant capacity via spectrophotometry. Data are expressed as mean ± SD. Statistical analysis was performed using one way ANOVA followed by Tukey’s multiple comparison test. (***) significance compared to the control group with p value < 0.05. (+ + +) significance of AMPE treated groups (G4, G5) compared to ulcerative colitis group (G3) with p value < 0.05. (@@) comparison of G5(200 mg/kg AMPE) Vs G4 (100 mg/kg AMPE) with p value < 0.05. ns means non-significant

Regarding the oxidative stress marker, malondialdehyde (MDA) level was significantly abrogated in AMPE-treated groups 4 and 5 compared to colitis group 3 (Fig. 4b). The total antioxidant capacity (TAC) was significantly diminished in the colitis group (G3), indicating impaired redox homeostasis (Fig. 4c). Treatment with APME led to a substantial restoration of TAC in both treatment groups, with G5 achieving levels comparable to or exceeding those of the control group (G1), indicating enhanced antioxidant defense mechanisms.

Our data showed that HO-1 expression levels were significantly diminished in G3 (colitis group) compared to the control group. Treatment with AMPE restored the HO-1 gene expression in the treated groups (G4 and G5) compared to the colitis group in a dose-dependent manner (Fig. 5f). The Keap1/Nrf2 signalling pathway, comprising Nrf2 and its inhibitory counterpart, Kelch-like ECH-associated protein 1 (Keap1), has been shown to exert a protective effect in both experimental models and individuals with ulcerative colitis (Fig. 5d, e).

Fig. 5.

Fig. 5

Open in a new tab

Quantitative PCR gene expression levels a FENDRR b Neat1 c NFκB d Nrf2 e Keap1 f HO-1. Data are expressed as mean ± SD. Statistical analysis was performed using one way ANOVA followed by Tukey’s multiple comparison test. (***) significance compared to the control group with p value < 0.05. (+ + +) significance of AMPE treated groups (G4, G5) compared to ulcerative colitis group (G3) with p value < 0.05. (@@) comparison of G5(200 mg/kg AMPE) Vs G4 (100 mg/kg AMPE) with p value < 0.05. ns means non-significant

Assessment of gene expression levels of FENDRR and Neat1 LncRNAs

The current findings demonstrate a marked upregulation of both FENDRR and Neat1 in G3, followed by a progressive downregulation in G4 and G5, suggesting a disease-associated elevation with subsequent modulation by treatment (Fig. 5a, b).

FENDRR expression

In our study, FENDRR was significantly elevated in G3 (18.44 ± 1.00), indicating strong transcriptional activation, likely in response to inflammatory or fibrotic signaling pathways. The observed mitigation of FENDRR levels in groups 4 and 5 (11.619 ± 1.2 and 7.25 ± 0.289, respectively) likely reflects the intervention’s capacity to reduce inflammation-mediated fibrotic gene expression.

NEAT1 expression

Similarly, NEAT1 expression peaked in GP 3 (8.3 ± 1.054), supporting its well-documented role in inflammation and immune regulation (Liu et al. 2018). Notably, the downregulation observed in GPs 4 and 5 (2.546 ± 0.362 and 1.6 ± 0.135, respectively) indicates a potential anti-inflammatory effect.

Discussion

AP is a halophytic perennial member of the genus Atriplex with restricted phytochemical and biological investigations (Zanella and Vianello 2020; Hamada et al. 2025). Despite Abd El Raheim (2013) reporting that pretreatment with 400 mg/kg extracts of Atriplex farinosa and or Atriplex nummularia effectively protected against colonic ulceration and attenuated the elevation of myeloperoxidase activity, no anti-UC study was reported at AP.

Bioactive phytoconstituents contribute significantly to the maintenance of human health and alleviate organ malfunction (Davila and Papada 2023). Antibacterial organic acid may support the intestinal barrier integrity (Raybaudi-Massilia et al. 2009; Ricciutelli et al. 2020; Mu et al. 2012; Fukahori et al. 1996).

Caffeic acid is a common hydroxycinnamic acid found in many fruits. It showed a significant role in modulating intestinal inflammation by reducing COX-2 expression, PGE2 production, and IL-8 biosynthesis in IL-1β-treated human colonic myofibroblasts, while also inhibiting advanced glycation end product formation through strong chelating activity (Zieli´nska et al. 2021).

Flavonoids possess considerable therapeutic value in the management of inflammatory diseases due to their ability to inhibit various pro-inflammatory proteins, thereby functioning as natural modulators and reducers of inflammation`s severity (García-Lafuente et al. 2009). Beyond their direct antioxidant activity, flavonoids also regulate the expression of numerous antioxidants and cytoprotective genes through modulation of nuclear transcription factors and suppression of key inflammatory pathways (Lu et al. 2013). Furthermore, flavonoids positively influence gut microbiota composition by promoting the growth of beneficial bacteria such as Bifidobacterium and Lactobacillus species, thus contributing to an anti-inflammatory intestinal environment (Duda-Chodak et al. 2015; Parkar et al. 2013; Cardona et al. 2013; Satokari 2020). According to Formiga (2020), the co-administration of rosmarinic acid and p-cymene demonstrated notable anti-inflammatory activity within the intestinal tract.

Meanwhile, the essential oil of Bunium persicum decreased acetic acid-induced rat colitis via suppression of the NF-κB pathway (Rashidian et al. 2021).

In the context of IBD treatments, stigmasterol exhibits notable anti-inflammatory effects by modulating gut microbiota, enhancing butyrate production, suppressing key cytokines via NF-κB inhibition, strengthening the mucosal immunity, and intestinal barrier function. Moreover, 20-hydroxyecdysone showed antioxidant and anti-inflammatory activities (Wen et al. 2021; Jie et al. 2022; Bakrim et al. 2022; Dinan et al. 2021).

Research indicates that excessive dietary sugar intake alters gut microbiota composition by increasing the relative abundance of Proteobacteria while reducing Bacteroidetes, a phylum known to contribute to endotoxin neutralization and maintenance of gut barrier integrity. This imbalance shifts the microbial profile toward a pro-inflammatory state and impairs the regulation of epithelial and mucosal immune functions. As a result, high sugar consumption can drive metabolic endotoxemia and low-grade systemic inflammation, ultimately contributing to metabolic dysfunction. These findings suggest that, beyond supplying surplus energy, high dietary sugar may exert multiple adverse effects on health through microbiota-mediated pathways (Khakh and North 2012). Despite this evidence, Zhang (2022) found that sucrose reduces inflammatory cytokine expression only at low doses.

Our data on AP`s total phenolic and flavonoid contents are in harmony with those reported by Zanella & Vianello (2020). Among the thirty-five tentatively detected APME`s phytoconstituents [Table S1], organic acids [D-(+)-malic acid 1, 2-isopropylmalic acid 2, D-3-Phenyllactic acid 3, p-hydroxybenzoic acid 4, 2,5-dihydroxybenzoic acid 5], phenolic acids [caffeic acid 6 & rosmarinic acid 7], flavonoid glycosides [myricetin 8, baicalein-7-O-glucuronide 9, kaempferol-3-O-α-L-rhamnoside 10, isorhamnetin-3-O-rutinoside 11, okanin-4’-O-glucoside 12, kaempferol-7-O-neohesperidoside 13, kaempferol-3-Glucuronide 14, 3,5, 7-trihydroxy-4’-methoxyflavone 15] procyanidin B2 16, gluconate 18, UDP-xylose 27 and γ-terpinene are likely the key contributors to its observed anti-UC efficacy. We propose that their synergistic interaction may offset the adverse impact of its carbohydrate content.

Furthermore, two effective anti- IBD metabolites [stigmasterol 1 and 20-hydroxyecdysone 2] were isolated from APME`s fractions. To date, there is no definitive evidence that stigmasterol has been previously isolated from AP, although 20-hydroxyecdysone has been reported from its roots (Ben Nejma et al. 2015). These findings provide a strong incentive for exploring APME`s effectiveness in UC therapy through an in vivo study.

During our study of the survival group G2, it is reasonable to state that there is no evidence of AP`s toxicity as documented by Hamada (2025) and Abd El Raheim (2013).

Histological assessment of hematoxylin and eosin (H&E)-stained sections from UC`s patients showed a marked increase in the apoptosis rate of colonic mucosal epithelial cells. This heightened cell turnover, characterized by excessive apoptosis and compensatory proliferation, contributes to the structural disruption of crypt architecture and compromises the integrity of the intestinal mucosal barrier (Zhao et al. 2016). In renal fibrosis, Huangqi decoction demonstrated a dose-dependent protective effect on the ipsilateral kidney by suppressing the expression of TGF-β1, TGF-β receptors I and II, Smad2, phosphorylated Smad2 (P-Smad2), Smad4, α-SMA, and collagens I, III, and IV. Conversely, it enhanced the expression of the inhibitory regulator Smad7 (Luo et al. 2022). Likewise, our histological analysis of the tested groups confirmed APME's safety, ability to reduce induced inflammation and cytoprotective efficacy (Fig. 2A–F).

Colitis induction resulted in a marked elevation of proinflammatory mediators, including TNF-α, PTX3, and PAF in plasma, as well as increased TLR4 expression in colonic tissue (Ercan et al. 2025). Phytochemicals capable of downregulating TNF-α secretion or interfering with its associated inflammatory signalling pathways may serve as promising alternatives for the management of immune-mediated inflammatory disorders (Subedi et al. 2020).

Overexpression of the p53 protein in colorectal crypts is frequently observed in ulcerative colitis (UC) patients, even in the absence of histopathological evidence of dysplasia. This overexpression is often interpreted by pathologists as an intermediate state between regenerative epithelial changes and intraepithelial neoplasia, and it serves as a valuable biomarker for assessing the risk of malignant transformation (Popp et al. 2016; Rubin and Turner 2006). Notably, a high frequency of p53 mutations has been reported in individuals with long-standing, severe UC who have not yet developed colorectal cancer, highlighting the role of chronic inflammation in driving early molecular alterations associated with carcinogenesis (Itzkowitz 2003; Hussain et al. 2000).

In the study conducted by Sukumari (2011), certain clinically relevant dietary phytochemicals significantly suppressed the proliferation of human S-type neuroblastoma (NB) cells. Notably, these antiproliferative effects appeared to be functionally independent of both caspase and p53 activation and were instead associated with inhibition of the Akt-NFκB signalling pathway (Sukumari-Ramesh et al. 2011).

Although COX-2 expression is generally low in the GI of healthy humans and animals, it plays a pivotal role in maintaining mucosal defence mechanisms. COX-2 is essential for promoting the resolution of inflammation, facilitating ulcer healing, and mediating long-term adaptive changes in GI function following inflammatory episodes (Wallace and Devchand 2005). The phytochemicals identified by Desai (2018) have demonstrated significant inhibitory effects on cyclooxygenase-2 (COX-2), an enzyme commonly upregulated during inflammatory processes and tumorigenesis through cancer-associated signalling pathways (Desai et al. 2018). Similarly, combined administration of both methanolic extracts from Egyptian propolis and Moringa oleifera Lam. (Moringaceae) exhibited a protective effect against acetic acid-induced ulcerative colitis in rats. Both synergistically inhibited COX-1 and COX-2 activity, lowered the ulcerative index and lesion scores, and attenuated oxidative stress markers and pro-inflammatory mediators (Atta et al. 2019). These findings are consistent with previous studies demonstrating that polyphenol and flavonoid rich plant extracts reduce COX-2 expression and inflammatory mediators in chemically induced colitis models (Salaritabar et al. 2017). The ability of APME to diminish COX-2 overexpression reinforces its therapeutic value in managing UC through modulation of inflammatory enzyme pathways.

Importantly, APME remedies noticed markedly reduced TNF-α, p53, and Cox-2 expression levels in a dose-dependent manner (Fig. 3(1, 2 & 3(A–F))).

Lipid peroxidation, reflected by MDA levels, was significantly attenuated in AMPE-treated groups 4 and 5 compared to colitis group 3 (Fig. 4b). This indicates the efficacy of AMPE in the amelioration of oxidative stress induced by inflammation. On the other hand, total antioxidant capacity (TAC) was significantly diminished in the colitis group (G3), indicating impaired redox homeostasis (Fig. 4c). Treatment with APME led to a substantial restoration of TAC in both treatment groups, with G5 achieving levels comparable to or exceeding those of the control group (G1), indicating enhanced antioxidant defense mechanisms. These line with that nuclear factor erythroid 2–related factor 2 (Nrf2) serves as a key transcriptional regulator responsible for maintaining redox balance and activating cellular antioxidant defences (He et al. 2020).

The upregulated levels of TNF-α in the colitis group resulted from the deregulation of NFκB signalling induced by concomitant oxidative stress (Wang et al. 2016). Our data revealed significant upregulation of NFκB gene expression in the G3 colitis group, which is significantly counteracted by treatment with AMPE in treated groups (G4, G5) in a dose-dependent manner (Fig. 5c). In the same context, Zahedipour (2022) highlighted the downregulating effect of several phytochemicals on TNF-α in inflammatory conditions.

Hemeoxygenase-1 (HO-1) is a downstream target of Nr2 (Alsharif et al. 2022). Our data showed that HO-1 expression levels were significantly diminished in G3 (colitis group) compared to the control group. Treatment with AMPE restored the HO-1 gene expression in the treated groups (G4 and G5) compared to the colitis group in a dose-dependent manner (Fig. 5f). The Keap1/Nrf2 signalling pathway, comprising Nrf2 and its inhibitory counterpart, Kelch-like ECH-associated protein 1 (Keap1), has been shown to exert a protective effect in both experimental models and individuals with ulcerative colitis (Fig. 5d,e). This pathway is thus essential for the antioxidant response and intestinal protection (Piotrowska et al. 2021).

Collectively, these findings suggest that APME exerts potent anti-inflammatory effects by inhibiting TNF-α expression, while concurrently restoring antioxidant capacity in colonic tissues. This dual action supports APME's potential as a complementary therapeutic agent in managing ulcerative colitis by targeting both inflammatory cytokines and oxidative stress.

For instance, Adylova (2021) reported that FENDRR expression was markedly elevated in colonic tissues of mice with DSS-induced colitis and was associated with enhanced TGF-β/Smad signaling activity. Similarly, Sun and Kraus (2015) found FENDRR upregulation in models of cardiac and pulmonary fibrosis, supporting its role as a pro-fibrotic LncRNA. The decline in FENDRR expressions in GPs 4 and 5 after treatment may reflect the therapeutic effect of the intervention in suppressing inflammation-induced fibrotic gene expression, consistent with findings from Lee (2022) who observed that pharmacological inhibition or genetic knockdown of FENDRR reduced fibrosis and inflammation in experimental colitis models.

lncRNAs NEAT1 are involved in the underlying mechanisms of UC. Studies using IBD mouse models have demonstrated that suppression of NEAT1 expression attenuates the inflammatory response, enhances intestinal barrier function, and modulates macrophage polarization toward an anti-inflammatory phenotype. Elevated NEAT1 expression has been associated with impaired intestinal barrier integrity in IBD, contributing to disease progression and mucosal dysfunction. The elevated NEAT1 expression in our study mirrors this pattern, suggesting its contribution to colonic inflammation and tissue damage (Liu et al. 2018).

Liu (2018) documented that NEAT1 expression is significantly upregulated in a murine model of IBD, where it contributes to the modulation of intestinal inflammation through exosome-mediated regulation of macrophage polarization. Notably, silencing NEAT1 alleviated intestinal inflammation in mice; however, the precise molecular mechanisms underlying NEAT1’s role in intestinal immune regulation remain incompletely understood.

Overall, the expression trends of FENDRR and NEAT1 in this study are consistent with existing literature that links these lncRNAs to inflammatory and fibrotic responses in colonic and systemic disease models. The observed reduction in expression following treatment supports their role as biomarkers of disease activity and potential targets for therapeutic modulation.

Conclusion

AP is a halophytic plant with variant bioactive phytochemicals (Zanella and Vianello 2020), yet its anti-UC impact remains unexplored in animal models. APME was assessed for its total phenolic and flavonoid contents. Negative ionization mode LC–ESI–MS/MS analysis of APME revealed 35 bioactive metabolites. Additionally, stigmasterol 1 and 20-hydroxyecdysone 2 were isolated from the n-hexane and ethyl acetate fractions, respectively, using column chromatography followed by spectroscopic characterization. APME demonstrates promising antioxidant and anti-inflammatory properties in a dose-responsive pattern. It ameliorates histological damage, reduces inflammatory cytokine production, and restores oxidative balance. These effects are mediated by modulation of transcription factors (Nrf2, NF-κB), inflammatory mediators (TNF-α, COX-2, P53), and regulatory lncRNAs (FENDRR, Neat1). Although these findings support the therapeutic potential of APME, further investigations are warranted, including clinical trials to establish its efficacy.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1. (118.1KB, docx)

Author contributions

M.H., M.El-A., A.K., G.A. Conceptualization, M.H., M.M.M., M.M.A., M.El-A., A.K., G.A. Methodology, M.H., M.M.M., M.M.A., M.El-A., A.K., G.A. Investigation, M.H., M.M.M., M.M.A., M.El-A., A.K., G.A. Formal analysis, M.H., M.M.M., M.M.A., M.El-A., A.K., G.A. Data curation, M.M.M., M.M.A., M.El-A., A.K., G.A. Writing—original draft., M.El-A., A.K., G.A. Supervision. Finally, M.El-A., A.K., M.M.M., M.M.A., G.A. Visualization, Writing—Review & Editing.

Funding

Open access funding provided by The Science, Technology & Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB). This study was self-funded.

Data availability

All data generated or analyzed during this study are included in this published article and its supplementary information files. Additional details are available from the corresponding author upon reasonable request.

Declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. Abd El Raheim DM, Soliman GA, Khataibeh MH, Alqasoumi SI, Ansari MN (2013) Effect of Atriplex farinose, Atriplex nummularia and Ficus ingens on ulcerative colitis in rats. IJBPAS 2(6):1247–1259 [Google Scholar]
  2. Abdelgawad FAM, El-Hawary SS, Abd El-Kader EM (2024) Phytochemical elucidation and effect of Maesa indica (Roxb.) sweet on alleviation of potassium dichromate-induced pulmonary damage in rats. Plants 13(3):338. 10.3390/plants13030338 [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Abdl Aziz FT, Temraz A, Hassan MA (2024) Metabolites profiling by LC-ESI-MS/MS technique and in-vitro antioxidant activity of Bauhinia madagascariensis Desv. and Bauhinia purpurea L. aerial parts cultivated in Egypt: a comparative study. Azhar Int J Pharm Med Sci 4(1):169–188. 10.21608/aijpms.2023.212409.1215 [DOI] [Google Scholar]
  4. Abdulla M, Mohammed NA (2022) Review on inflammatory bowel diseases: Recent molecular pathophysiology advances. Biol Targets Ther 16:129–140. 10.2147/BTT.S380027 [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Adylova A, Mukhanbetzhanovna AA, Attar R, Yulaevna IM, Farooqi AA (2021) Regulation of TGFβ/SMAD signaling by long non-coding RNAs in different cancers: Dark Knight in the Castle of molecular oncology. Noncoding RNA Res 6(1):23–28. 10.1016/j.ncrna.2020.12.003.PMID:33511320;PMCID:PMC7814108 [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Alsharif IA, Fayed HM, Abdel-Rahman RF, Abd-Elsalam RM, Ogaly HA (2022) Miconazole mitigates acetic acid-induced experimental colitis in rats: insight into inflammation, oxidative stress, and Keap1/Nrf-2 signaling crosstalk. Biology (Basel) 11(2):303. 10.3390/biology11020303 [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Atta AH, Mouneir SM, Nasr SM, Sedky D, Mohamed AM, Atta SA, Desouky HM (2019) Phytochemical studies and anti-ulcerative colitis effect of Moringa oleifera seeds and Egyptian propolis methanol extracts in a rat model. Asian Pac J Trop Biomed 9(3):98–108. 10.4103/2221-1691.254603 [DOI] [Google Scholar]
  8. Attiq A, Jalil J, Husain K, Ahmad W (2018) Raging the war against inflammation with natural products. Front Pharmacol 9(976):1–27 [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bakrim S, Benkhaira N, Bourais I, Benali T, Lee L-H, El Omari N, Sheikh RA, Goh KW, Ming LC, Bouyahya A (2022) Health benefits and pharmacological properties of stigmasterol. Antioxidants 11(10):1912. 10.3390/antiox11101912 [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bancroft JD, Layton C (2010) The hematoxylins and eosin. Bancroft’s theory and practice of histological techniques, 7th ed. Churchill Livingstone Elsevier, Oxford, UK [Google Scholar]
  11. Ben Nejma A, Nguir A, Jannet HB, Hamza MA, Daïch A, Othman M, Lawson AM (2015) New septanoside and 20-hydroxyecdysone septanoside derivative from Atriplex portulacoides roots with preliminary biological activities. Bioorg Med Chem Lett 25(8):1665–1670. 10.1016/j.bmcl.2015.03.028 [DOI] [PubMed] [Google Scholar]
  12. Cardona F, Andrés-Lacueva C, Tulipani S, Tinahones FJ, Queipo-Ortuño MI (2013) Benefits of polyphenols on gut microbiota and implications in human health. J Nutr Biochem 24(8):1415–1422. 10.1016/j.jnutbio.2013.05.001 [DOI] [PubMed] [Google Scholar]
  13. Collins D (2022) A HPLC-ESI-MS/MS study of hydroxybenzoic acids and related derivatives in commercial seaweed biostimulants and their plant growth bioactivity. Victoria University, Melbourne, Australia, Institute for sustainable industries and livable cities [Google Scholar]
  14. Courtois G, Gilmore TD (2006) Mutations in the NF-κ B signaling pathway: implications for human disease. Oncogene 25(51):6831–6843. 10.1038/sj.onc.1209939 [DOI] [PubMed] [Google Scholar]
  15. Davila MM, Papada E (2023) The role of plant-derived natural products in the management of inflammatory bowel disease—What is the clinical evidence so far? Life (Basel) 13(8):1703. 10.3390/life13081703 [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Desai SJ, Prickril B, Rasooly A (2018) Mechanisms of phytonutrient modulation of cyclooxygenase-2 (COX-2) and inflammation related to cancer. Nutr Cancer 70(3):350–375. 10.1080/01635581.2018.1446091 [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dinan L, Dioh W, Veillet S, Lafont R (2021) 20-Hydroxyecdysone, from plant extracts to clinical use: therapeutic potential for the treatment of neuromuscular, cardio-metabolic and respiratory diseases. Biomedicines 9(5):492. 10.3390/biomedicines9050492 [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Duda-Chodak A, Tarko T, Satora P, Sroka P (2015) Interaction of dietary compounds, especially polyphenols, with the intestinal microbiota: a review. Eur J Nutr 54(3):325–341. 10.1007/s00394-015-0852-y [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. El-Zawahry HME, Shakour ZTA, El-Shafei AA, El-tantawy ME (2025) Larvicidal and cytotoxic activity of Cordyline terminalis Kunth and its metabolites profiling via UPLC-MS/MS. Azhar Int J Pharm Med Sci 5(1):141–151. 10.21608/aijpms.2024.256016.1246 [DOI] [Google Scholar]
  20. Ercan G, Aygun H, Akbaş A, Çınaroğlu OS, Erbas O (2025) Indoximod attenuates inflammatory responses in acetic acid-induced acute colitis by modulating Toll-like receptor 4 (TLR4) signaling and proinflammatory cytokines in rats. Medicina (b Aires) 61(6):1033. 10.3390/medicina61061033 [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fabre N, Isabelle R, de Hoffmann E, Quetin-Leclercq J (2001) Determination of flavone, flavonol, and flavanone aglycones by negative ion liquid chromatography electrospray ion trap mass spectrometry. J Am Soc Mass Spectrom 12(6):707–715. 10.1016/S1044-0305(01)00226-4 [DOI] [PubMed] [Google Scholar]
  22. Formiga RO, Alves Júnior EB, Vasconcelos RC, Guerra GCB, Antunes de Araújo A, deCarvalhoTG, Garcia VB, deAraújo, Junior RF, Gadelha FAAF, Vieira GC, Sobral MV, Filho JMB, Spiller F, Batista LM (2020) p-Cymene and rosmarinic acid ameliorate TNBS-induced intestinal inflammation upkeeping ZO-1 and MUC-2. Int J Mol Sci 21(16):5870. 10.3390/ijms21165870. [DOI] [PMC free article] [PubMed]
  23. Fukahori M, Akatsu S, Sato H, Yotsuyanagi T (1996) Relationship between uptake of p-hydroxybenzoic acid esters by Escherichia coli and antibacterial activity. Chem Pharm Bull 44(8):1567–1570. 10.1248/cpb.44.1567 [DOI] [PubMed] [Google Scholar]
  24. García-Lafuente A, Guillamón E, Villares A, Rostagno MA, Martínez JA (2009) Flavonoids as anti-inflammatory agents: implications in cancer and cardiovascular disease. Inflamm Res 58(9):537–552. 10.1007/s00011-009-0037-3 [DOI] [PubMed] [Google Scholar]
  25. Gerondakis S, Grossmann M, Nakamura Y, Pohl T, Grumont R (1999) Genetic approaches in mice to understand Rel/NF-κB and IκB function: transgenics and knockouts. Oncogene 18(49):6888–6895. 10.1038/sj.onc.1203236 [DOI] [PubMed] [Google Scholar]
  26. Hamada MS, Abd-El-Aziz MH, Alsenosy NK, Eltelet DH (2025) Evaluation of anti-cytogenotoxic potential of Atriplex portulacoides crude extract in mice bone marrow. DJAS 4(II):116–122. 10.21608/djas.2025.429289 [DOI] [Google Scholar]
  27. Hazel K, O’Connor A (2020) Emerging treatments for inflammatory bowel disease. Ther Adv Chronic Dis 11:1–12. 10.1177/2040622319899297 [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. He W, Zhong G, Wang P, Jiang C, Jiang N, Huang J (2019) Downregulation of long noncoding RNA FENDRR predicts poor prognosis in renal cell carcinoma. Oncol Lett 17(1):103–112. 10.3892/ol.2018.9624 [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. He F, Ru X, Wen T (2020) NRF2, a transcription factor for stress response and beyond. Int J Mol Sci 21(13):4777. 10.3390/ijms21134777 [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hussain SP, Amstad P, Raja K, Ambs S, Nagashima M, Bennet WP, Shields PG, Ham AJ, Swenberg JA, Marrogi AJ, Harris CC (2000) Increased p53 mutation load in noncancerous colon tissue from ulcerative colitis: a cancer-prone chronic inflammatory disease. Cancer Res 60(13):3333–3337 [PubMed] [Google Scholar]
  31. Itzkowitz S (2003) Colon carcinogenesis in inflammatory bowel disease: applying molecular genetics to clinical practice. J Clin Gastroenterol 36(5S):S70–S74. 10.1097/00004836-200305001-00012 [DOI] [PubMed] [Google Scholar]
  32. Jie F, Yang X, Yang B, Liu Y, Wu L, Lu B (2022) Stigmasterol attenuates inflammatory response of microglia via NF-κB and NLRP3 signaling by AMPK activation. Biomed Pharmacother 153:113317 [DOI] [PubMed] [Google Scholar]
  33. Khakh BS, North RA (2012) Neuromodulation by extracellular ATP and P2X receptors in the CNS. Neuron 76(1):51–69. 10.1016/j.neuron.2012.09.024 [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kubiak-Martens L, Brinkkemper O, Oudemans TFM (2015) What’s for dinner? Processed food in the coastal area of the northern Netherlands in the Late Neolithic. Veg Hist Archaeobot 24(1):47–62. 10.1007/s00334-014-0485-8 [DOI] [Google Scholar]
  35. Lee HJ (2022) Therapeutic potential of the combination of pentoxifylline and vitamin E in inflammatory bowel disease: Inhibition of intestinal fibrosis. J Clin Med 11(16):4713. 10.3390/jcm11164713.PMID:36012952;PMCID:PMC9410449 [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Li H-Y, Wang S-M, Liu S-M, Bu S-S, Li J, Han D, Zhang M-z, Wu G-Y (2009) Separation and identification of purine nucleosides in the urine of patients with cancer by reverse phase liquid chromatography/electrospray tandem mass spectrometry. J Mass Spectrom 44(5):641–651 [DOI] [PubMed] [Google Scholar]
  37. Li F, Zhang J, Arfuso F, Chinnathambi A, Zayed ME, Alharbi SA, Kumar AP, Ahn KS, Sethi G (2015) NF-κB in cancer therapy. Arch Toxicol 89(5):711–731. 10.1007/s00204-015-1470-4 [DOI] [PubMed] [Google Scholar]
  38. Liu R, Tang A, Wang X, Chen X, Zhao L, Xiao Z, Shen S (2018) Inhibition of lncRNA NEAT1 suppresses the inflammatory response in IBD by modulating the intestinal epithelial barrier and by exosome-mediated polarization of macrophages. Int J Mol Med 42(5):2903–2913. 10.3892/ijmm.2018.3829 [DOI] [PubMed] [Google Scholar]
  39. Lu MF, Xiao ZT, Zhang HY (2013) Where do health benefits of flavonoids come from? Biochem Biophys Res Commun 434(4):701–704. 10.1016/j.bbrc.2013.04.035 [DOI] [PubMed] [Google Scholar]
  40. Luo Q, Huang S, Zhao L, Liu J, Ma Q, Wang Y, Dong Y, Li C, Qiu P (2022) Chang qing formula ameliorates colitis-associated colorectal cancer via suppressing IL17/NF-κB/STAT3 pathway in mice as revealed by network pharmacology study. Front Pharmacol 3(13):893231. 10.3389/fphar.2022.893231 [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Mattson MP, Meffert MK (2006) Roles for NF-κB in nerve cell survival, plasticity, and disease. Cell Death Differ 13(5):852–860. 10.1038/sj.cdd.4401837 [DOI] [PubMed] [Google Scholar]
  42. Millar AD, Rampton DS, Chander CL, Claxson AW, Blades S, Coumbe A, Panetta J, Morris CJ, Blake DR (1996) Evaluating the antioxidant potential of new treatments for inflammatory bowel disease using a rat model of colitis. Gut 39(3):407–415. 10.1136/gut.39.3.407 [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Mu W, Yu S, Zhu L, Zhang T, Jiang B (2012) Recent research on 3-phenyllactic acid, a broad-spectrum antimicrobial compound. Appl Microbiol Biotechnol 95(5):1155–1163. 10.1007/s00253-012-4269-8 [DOI] [PubMed] [Google Scholar]
  44. Oeckinghaus A, Hayden MS, Ghosh S (2011) Crosstalk in NF-κB signaling pathways. Nat Immunol 12(8):695–698. 10.1038/ni.2065 [DOI] [PubMed] [Google Scholar]
  45. Owusu G, Obiri DD, Ainooson GK, Osafo N, Antwi AO, Duduyemi BM, Ansah C (2020) Acetic acid-induced ulcerative colitis in Sprague Dawley rats is suppressed by the hydroethanolic extract of Cordia vignei leaves through reduced serum levels of TNF-α and IL-6. Int J Chronic Dis. 10.1155/2020/8785497 [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Parkar SG, Trower TM, Stevenson DE (2013) Fecal microbial metabolism of polyphenols and its effects on human gut microbiota. Anaerobe 23:12–19. 10.1016/j.anaerobe.2013.07.009 [DOI] [PubMed] [Google Scholar]
  47. Piotrowska M, Swierczynski M, Fichna J, Piechota-Polanczyk A (2021) The Nrf2 in the pathophysiology of the intestine: molecular mechanisms and therapeutic implications for inflammatory bowel diseases. Pharmacol Res 163:105243. 10.1016/j.phrs.2020.105243 [DOI] [PubMed] [Google Scholar]
  48. Popp C, Nichita L, Voiosu T, Bastian A, Cioplea M, Micu G, Pop G, Sticlaru L, Bengus A, Voiosu A, Mateescu RB (2016) Expression profile of p53 and p21 in large bowel mucosa as biomarkers of inflammatory-related carcinogenesis in ulcerative colitis. Dis Markers 2016:1–7. 10.1155/2016/3625279 [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Rashidian A, Akbarzadeh D, Asgarpanah J, Dehpour A (2021) Bunium persicum essential oil reduced acetic acid-induced rat colitis through suppression of NF-κB pathway. AJP 11(5):505–514. 10.22038/AJP.2021.18037 [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Raybaudi-Massilia RM, Mosqueda-Melgar J, Martín-Belloso O (2009) Antimicrobial activity of malic acid against Listeria monocytogenes, Salmonella enteritidis and Escherichia coli O157: H7 in apple, pear and melon juices. Food Control 20(2):105–112. 10.1016/j.foodcont.2008.02.009 [DOI] [Google Scholar]
  51. Ricciutelli M, Bartolucci G, Campana R, Salucci S (2020) Quantification of 2-and 3-isopropylmalic acids in forty Italian wines by UHPLC-MS/MS triple quadrupole and evaluation of their antimicrobial, antioxidant activities and biocompatibility. Food Chem 321(5):126726. 10.1016/j.foodchem.2020.126726 [DOI] [PubMed] [Google Scholar]
  52. Rubin DT, Turner JR (2006) Surveillance of dysplasia in inflammatory bowel disease: the gastroenterologist-pathologist partnership. Clin Gastroenterol Hepatol 4(11):1309–1313. 10.1016/j.cgh.2006.09.010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Ryhage R, Sydow E (1963) Mass spectrometry of terpenes. Acta Chem Scand 17(7):2025–2035 [Google Scholar]
  54. Salaritabar A, Darvishi B, Hadjiakhoondi F, Manayi A, Sureda A, Nabavi SF, Fitzpatrick LR, Nabavi SM, Bishayee A (2017) Therapeutic potential of flavonoids in inflammatory bowel disease: a comprehensive review. World J Gastroenterol 23(28):5097–5114. 10.3748/wjg.v23.i28.5097 [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Satokari R (2020) High intake of sugar and the balance between pro- and anti-inflammatory gut bacteria. Nutrients 12(5):1348. 10.3390/nu12051348 [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Sri Harsha PSC, Abdul Wahab R, Cuparencu C, Dragsted LO, Brennan LA (2018) Metabolomics approach to the identification of urinary biomarkers of pea intake. Nutrients 10(12):1911. 10.3390/nu10121911 [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stanojević L, Zvezdanović J, Danilović B, Cvetković D, Stanojević J, Ilić D, Milorad C (2018) The antioxidative and antimicrobial activity of the aqueous earth smoke (Fumaria officinalis L.) extract. Adv Technol 7(2):31–40 [Google Scholar]
  58. Subedi L, Lee SE, Madiha S, Gaire BP, Jin M, Yumnam S, Kim SY (2020) Phytochemicals against TNFα-mediated neuroinflammatory diseases. Int J Mol Sci 21(3):764. 10.3390/ijms21030764 [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Sukumari-Ramesh S, Bentley JN, Laird MD, Singh N, Vender JR, Dhandapani KM (2011) Dietary phytochemicals induce p53- and caspase-independent cell death in human neuroblastoma cells. Int J Dev Neurosci 29(7):701–710. 10.1016/j.ijdevneu.2011.06.002 [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Sun M, Kraus WL (2015) From discovery to function: the expanding roles of long noncoding RNAs in physiology and disease. Endocr Rev 36(1):25–64. 10.1210/er.2014-1034 [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Taniguchi K, Karin M (2018) NF-κB, inflammation, immunity and cancer: coming of age. Nat Rev Immunol 18(5):309–324. 10.1038/nri.2017.142 [DOI] [PubMed] [Google Scholar]
  62. Taylora VF, Marchb RE, Longerichc HP, Stadeyb CJ (2005) A mass spectrometric study of glucose, sucrose, and fructose using an inductively coupled plasma and electrospray ionization. Int J Mass Spectrom 243(1):71–84. 10.1016/j.ijms.2005.01.001 [DOI] [Google Scholar]
  63. Vuyyuru SK, Kedia S, Sahu P, Ahuja V (2022) Immune-mediated inflammatory diseases of the gastrointestinal tract: beyond Crohn’s disease and ulcerative colitis. JGH Open 6(2):100–111. 10.1002/jgh3.12706 [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Wallace JL, Devchand PR (2005) Emerging roles for cyclooxygenase-2 in gastrointestinal mucosal defense. Br J Pharmacol 145(3):275–282. 10.1038/sj.bjp.0706201 [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Wang Z, Li S, Cao Y, Tian X, Zeng R, Liao D-F, Cao D (2016) Oxidative stress and carbonyl lesions in ulcerative colitis and associated colorectal cancer. Oxid Med Cell Longev 2016:9875298. 10.1155/2016/9875298 [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Wen S, He L, Zhong Z, Zhao R, Weng S, Mi H, Liu F (2021) Stigmasterol restores the balance of Treg/Th17 cells by activating the butyrate-PPARg axis in colitis. Front Immunol 12:741934. 10.3389/fimmu.2021.741934 [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Yuan Y, Hou W, Tang M, Luo H (2008) Separation of flavonoids from the leaves of Oroxylum indicum by HSCCC. Chromatographia 68(11):885–892. 10.1365/s10337-008-0859-0 [DOI] [Google Scholar]
  68. Zahedipour F, Hosseini SA, Henney NC, Barreto GE, Sahebkar A (2022) Phytochemicals as inhibitors of tumor necrosis factor alpha and neuroinflammatory responses in neurodegenerative diseases. Neural Regen Res 17(8):1675–1684. 10.4103/1673-5374.332128 [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Zanella L, Vianello F (2020) Functional food from endangered ecosystems: Atriplex portulacoides as a case study. Foods 9(11):1533. 10.3390/foods9111533 [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Zhang Y-Z, Li Y-Y (2014) Inflammatory bowel disease: Pathogenesis. World J Gastroenterol 20(1):91–99 [DOI] [PMC free article] [PubMed] [Google Scholar]
  71. Zhang X, Zhang B, Peng B, Wang J, Hu Y, Wang R, Wang S (2022) Different dose of sucrose consumption divergently influences gut microbiota and PPAR-γ/MAPK/NF-κB pathway in DSS-induced colitis mice. Nutrients 14(13):2765. 10.3390/nu14132765 [DOI] [PMC free article] [PubMed] [Google Scholar]
  72. Zhao J, Wang L, Cao A-L, Jiang M-Q, Chen X, Wang Y, Wang Y-M, Wang H, Zhang X-M, Peng W (2016) Huang Qi decoction ameliorates renal fibrosis via TGF-β/Smad signaling pathway in vivo and in vitro. Cell Physiol Biochem 38(5):1761–1774. 10.1159/000443115 [DOI] [PubMed] [Google Scholar]
  73. Zieli’nska D, Zieli’nski H, Laparra-Llopis JM, Szawara-Nowak D, Joanna Honke J, Giménez-Bastida JA (2021) Caffeic acid modulates processes associated with intestinal inflammation. Nutrients. 10.3390/nu13020554 [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Material 1. (118.1KB, docx)

Data Availability Statement

All data generated or analyzed during this study are included in this published article and its supplementary information files. Additional details are available from the corresponding author upon reasonable request.

Articles from Inflammopharmacology are provided here courtesy of Springer

RESOURCES